Novel protein and gene encoding the protein

ABSTRACT

A novel mechanosensitive channel protein derived from mouse or humans which is expressed specifically in the kidney and has the function of non-selectively transporting cation into cells in response to a mechanical stimulus; a DNA encoding these proteins; a method of screening cation channel activity activators or inhibitors with these proteins; and antibodies to these proteins. The above-described proteins are useful as a diagnostic agent or a therapeutic agent for diseases based on abnormalities in the cation channel, such as hypertension or diabetes, or as a tool for screening chemicals for preventing and/or treating these diseases.

FIELD OF THE INVENTION

[0001] The present invention relates to a novel mechanosensitive channel protein derived from mice or humans which is expressed specifically in the kidney (SAC1 (mouse) and hSAC (human)), DNA encoding said proteins, a method of screening cation channel activators or inhibitors using the proteins, and antibodies to the proteins.

[0002] The protein, DNA, and antibodies to the protein of the present invention are useful as a diagnostic agent or a therapeutic agent for diseases based on abnormalities in the cation channel such as hypertension or diabetes. They are also useful as a tool for screening chemicals for preventing and/or treating these diseases. In the present invention, mechanosensitive channel proteins such as SAC1, hSAC, and the like are generically referred to as SAC.

BACKGROUND ART

[0003] An ion channel is a protein with a pore structure which is located in biomembranes consisting of a lipid bilayer as a basic structure. The ion channel is classified according to the electrophysiological characteristics, i.e. gating, conductance, ion selectivity and the like. Ion channel molecules have multiple conformations. Namely, the ion channel molecules allow ions to permeate through in a certain state (open state), but do not allow ions to permeate in another state (closed state). Such a change in the conformation is referred to as opening and closing of a gate (gating). Three factors are known to control the gating: membrane potential, binding of low molecules (ligands) and membrane expansion.

[0004] There are many examples of voltage-dependent channels such as Na⁺ channel, Ca²⁺ channel, K⁺ channel, and the like. These channels have a molecular structure in which charges are gathered, called an electric sensor. Changes in the conformation are thought to occur by a force caused by an electric potential difference between the two sides the membrane. So ions can move through the pore. A ligand-sensitive channel is activated by binding of a specific ligand (agonist) from outside the cell. This is also called a receptor built-in channel. On the other hand, there is a channel whose opening and closing operations are controlled by binding of molecules with a low molecular weight, such as second messengers, existing inside the cells. An example is an IP₃ (inositol trisphosphate) receptor existing in cell membrane. A Ca²⁺ channel opens when IP₃ binds to the receptor at the cytoplasmic side. A mechanosensitive channel is an ion channel in which opening and closing of the gate is controlled by stretching of the membrane. Since most channels open the gate by stretching, these are also called “stretch sensitive” channels. The channel is thought to be involved in the osmotic pressure regulation and volume adjustment of cells. It is quite natural that the ion channel exists in a stretch receptor such as muscle spindles. The ion channels have been discovered in many kind of cells up to now (Morris C. E., J. Membrane Biol., 113, 93-107 (1990); Bear, C. E., Am. J. Physiol., 258, C421-8 (1990); Cemerik, D. & Sackin, H., Am. J. Physiol., 264, F697-714 (1993); Lansman, J. B., et al., Nature, 325, 811-3 (1987); Sackin, H., Am. J. Physiol., 253, F1253-62 (1987)).

[0005] In addition, it has been indicated that a K⁺ channel of mammals encoding a TWIK1 (tandem of P domains in a weak inward rectifier K⁺ channel) related TASK (TWIK-1 related acid-sensitive K⁺ channel) having four transmembrane domains possesses the characteristics of opening and closing by mechanical stimulus (Duprat, F. et al.,(1997) EMBO J., 16, 5464-71, Patel, A. J., et al.,(1998) EMBO J. 17, 4283-90).

[0006] The inventors of the present invention have conducted gene cloning based on the hypothesis that a vanilloid receptor sensitive to heat as a physical factor will be a mechanosensitive channel. As a result, the inventors have already presented a report on a cDNA similar to the vanilloid receptor (Caterina, M. J., et. al., (1997) Nature, 389,816-24) and encoding a nonselective cation channel (SIC) having an ankyrin repeat and six transmembrane domains and controlled by stretch (Suzuki, M., et. al., (1999) J. Biol. Chem. 274, 6330-5) Since a mechanosensitive channel can convert a mechanical stimulus into a Ca²⁺ influx, a stretch-activated (SA) nonselective cation channel is important.

DISCLOSURE OF THE INVENTION

[0007] In view of this situation, the inventors of the present invention have conducted an extensive investigation for a novel mechanosensitive channel protein. As a result, the inventors have found a protein expressed specifically in the kidney and having a function of non-selectively incorporating cations into cells in response to a mechanical stimulus, Namely a novel mechanosensitive channel protein. Accordingly, an object of the present invention is to provide a novel mechanosensitive channel protein derived from mice or humans expressed specifically in the kidney and possessing a function of non-selectively incorporating cations into cells in response to a mechanical stimulus (SAC1 (mouse) (Stretch Activated Channel protein 1) and hSAC (human) (human Stretch Activated Channel protein)), DNA encoding these proteins, a method of screening a cation channel activators or inhibitors using the proteins, and antibodies to the proteins.

[0008] The present invention relates to a novel mechanosensitive channel protein derived from mice or humans (SAC1 (mouse) and hSAC (human)) which is expressed specifically in the kidney and has a function of non-selectively incorporating cations into cells in response to a mechanical stimulus. The present invention also relates to DNA encoding the proteins. Furthermore, the present invention relates to a method of screening cation channel activators or inhibitors using these proteins. The present invention also relates to antibodies to these proteins. The proteins, the DNA, and the antibodies to the proteins can be used as a diagnostic agent or a therapeutic agent for diseases based on abnormalities in the cation channel, such as hypertension or diabetes, or as a tool for screening chemicals for preventing and/or treating these diseases.

[0009] Advanced technologies such as molecular biotechnology, cell biological technology, and electrophysiological technology are required in order to carry out the isolation, identification, and functional analysis of a novel membrane channel. The present inventors have isolated a cDNA of an SA cation channel (SAC1) from mouse kidney and have determined the amino acid sequence using these technologies. The cDNA belongs to a superfamily of vanilloid receptors and is localized in the uriniferous tubules of the kidney. The SAC1 was subjected to expression in a mammal cell line. Furthermore, a channel gene corresponding to that of humans was obtained using the SAC1 gene which is of mouse origin.

[0010] As a result of electrophysiological analysis, intracellular free calcium has been increased by mechanically stimulating transformed cells that express SAC1. This is proved to be a single channel activated by negative pressure, being blocked by Gd₃ ³⁺ and non-selective with respect to permeation of cations; the ionic permeability of!SAC1 is of Na⁺>K⁺. The reversal potential of the whole cells shifted toward positive by inflating the cells. In addition, since a deletion mutant of the channel becomes unresponsive to stretch, it is suggested that the SAC1 may become an SA channel.

[0011] As mentioned above, the present invention relates to a novel mechanosensitive channel protein derived from mice or humans (SAC1 (mouse) and hSAC (human)) which is expressed specifically in kidney and has a function of non-selectively incorporating cations into cells in response to a mechanical stimulus, and to DNA encoding these proteins. The protein of the present invention can be obtained as follows. A mouse kidney cDNA library is constructed and amplified with a PCR method using suitable primers to obtain SAC1 cDNA. hSAC cDNA can then be obtained by cloning from a human kidney cDNA library using this SAC1 cDNA as a probe. The SAC1 derived from mice is a protein having 871 amino acid residues, whereas the hSAC derived from humans is also a protein having 871 amino acid residues. These exhibit 94.4% homology between'the amino acid sequences thereof. This significant homology can be thought to indicate that SAC1 has common structures and functions in mammalian cells. The DNA encoding the hSAC has a length of about 18 Kb, is composed of 14 exons, and exists on chromosome 12q 24.1.

[0012] A mouse or human SAC protein can be obtained by inserting the obtained SAC gene derived from mouse or human in a suitable vector and-transforming the host cells with the vector. As host cells, bacteria, yeast, animal cells, and the like can be used. Particularly, HeLa cells, Chinese hamster ovary (CHO) cells, or COS-7 cells are preferable as animal cells. In addition, a promoter such as a virus polyoma, adenovirus, cytomegalovirus, or simian virus 40 can be used for controlling an expression plasmid of cell line. Particularly, as a useful plasmid in expression system of animal cells, pCMV is preferably known (Thomsen, et al., PNAS, (1984) 81, 659). The resulting protein is a membrane-bound protein sensitive to mechanical stimulus and can be used as a diagnostic agent or a therapeutic agent for diseases based on abnormalities in the cation channel such as hypertension or diabetes, also used as a tool for screening chemicals for preventing and/or treating these diseases.

[0013] In addition, the proteins of the present invention or the fragments thereof are useful for diagnosis of defective hSAC by DNA hybridization. Mutants of hSAC are useful for research of hypertension and diabetes. Furthermore, fusion proteins can be easily prepared by connecting a nucleotide sequence encoding other proteins or synthetic polypeptides to 5′ or 3′ terminals of SAC DNA or its mutant using conventionally known technology. For example, the fusion protein may be prepared as a precursor protein, and caused to function when digested in vitro or in vivo, and possess selective distribution in a objective tissue and the like in addition to the inherent functions.

[0014] Moreover, the expressed proteins, mutants, or fragments thereof may immunize an animal to produce a polyclonal antibody. In addition, a monoclonal antibody can be prepared using hybridoma cells produced by fusion of lymphocytes obtained from an immunized animal and myeloma cells.

[0015] The proteins of the present invention or their mutants or analogues can be used as a diagnostic agent or therapeutic agent for diseases relating to channel proteins, or also for screening substances agonistically or antagonistically acting on SAC. Acquisition of the DNA sequence information on SAC has made it easy to prepare a partial DNA or RNA sequence. Since such a partial DNA sequence is capable of hybridizing to genes to be selected, the DNA can be used as a nucleic acid probe. Such a probe is useful for detecting cDNA sequences in various tissues. It is possible to obtain nucleic acids able to hybridize from various organisms and their tissues using the probe prepared by using the SAC. The resulting nucleic acids include a nucleic acid encoding a protein exhibiting a same isotype as SAC or a novel feature. In addition, the probe prepared can be used for gene diagnosis of diseases. It is possible to detect a disease gene by identifying the nucleotide sequence from a patient hybridized with the probe. A gene therapeutic agent used for substantial treatment can also be prepared. The nucleotide sequence of SAC and its mutant or derivatives thereof can be incorporated in a plasmid or stem cells and administered as an agent for gene therapy.

[0016] The protein of the present invention and the antibodies to the protein can be administered safely to human being and primate animals. The protein of the present invention can be prepared to a pharmaceutically acceptable formula and administered orally or non-orally. Example pharmaceutical compositions include compositions for injection, infusion, suppositories, nasal agents, buccals, percutaneous absorption agents, and the like. These compositions are formulated according to known pharmaceutical preparation methods using pharmaceutically acceptable carriers, vehicles, stabilizers, coloring agents, surfactants, and/or other additives, and made into objective formulations. In preparing compositions for injection, a pharmacologically effective amount of protein of the present invention may be mixed with a pharmaceutically acceptable vehicles, such as amino acids, saccharides, cellulose derivatives, other organic compounds and/or inorganic compounds.

[0017] In addition, in preparing a composition for injection using the antibodies of the present invention such vehicles and/or activating agents, a pH adjusting agent, buffering agent, stabilizer, solubilizing agent, and the like, may be optionally added according to conventional methods.

[0018] The DNA of the present invention can be used for gene therapy either by itself alone, by incorporating into a liposome, or by inserting into a vector for gene therapy such as retrovirus and adenovirus.

BRIEF DESCRIPTION OF THE DRAWINGS

[0019]FIG. 1(a) shows results of northern blotting analyses of SAC1 mRNA expression using the full length cDNA. RNA (2 μg) prepared from mouse tissues was applied to each lane.

[0020]FIG. 1(b) shows a result of immunostaining of a kidney tissue. Stained areas are shown in white.

[0021]FIG. 2 shows a result of the effect of SAC1 expression cells (GFP positive) on touch stress. In this figure, ∘ indicates the fluorescence intensity at 380 nm, ▴ indicates the fluorescence intensity ratio (360 nm/380 nm), and  indicates the fluorescence intensity at 360 nm.

[0022]FIG. 3 shows results of response to stretch of SAC single channel, wherein

[0023] (A) shows a typical result of SAC1 channel when negative pressure was applied stepwise,

[0024] (B) shows the result when negative pressure was gradually applied, and

[0025] (C) shows the relationship between pressure and Po when the pressure was applied to SAC1 with or without addition of GdCl₃. In the Figure, ∘ indicates a control (pressure without the addition of GdCl₃) and Δ indicates pressure with the addition of 0.5 M GdCl₃, respectively.

PREFERRED EMBODIMENT OF THE INVENTION EXAMPLES

[0026] The present invention will now be described in more detail by way of examples, which are given for the purpose of explanation and should not be construed as limiting the present invention.

Example 1

[0027] Isolation of cDNA Encoding SAC1

[0028] RNA was isolated by the guanidine thiocyanate method using Trizol (Gibco-BRL). mRNA was purified using a polyA column (Pharmacia) The cDNA-library of mouse kidney was prepared using a Marathon cDNA construction kit (Clonetech Co.) The cDNA was amplified by PCR (30 cycles, one cycle consisting of 30 seconds at 94° C. and 4 minutes at 70° C.) using a primer set 1 (AA13f2 (SEQ ID No: 5 in Sequence Listing) and mA13end1 (SEQ ID No: 6)) and ExTaq polymerase (Takara, Ltd.). A nested primer (AA13r2 (SEQ ID No: 7)) was synthesized to determine the 5′ terminal by the 5′ PACE method.

[0029] Using the primer set 2 (A13st1 (SEQ ID No: 8) and AA13r1 (SEQ ID No: 9)) and ExTaq polymerase (Takara, Ltd.), PCR amplification was carried out under the same conditions as described above. Using each fragment obtained by PCR amplification using the two primer sets, the region containing the full length was amplified by PCR using primer set (A13st1 and mA13end1). SAC1 cDNA with a length of about 3.2 Kb was again cloned from the cDNA library. The cloned cDNA was ligated to a TA cloning vector (TOPO-XL; InVitrogen) and its BamHI-HindIII fragment was ligated to a mammalian cells expression vector (pCMV-SPORT; Gibco-BRL). Sequencing was carried out by an automatic sequencer (model 373-S; Applied Bioinstruments Inc.) using Thermo Sequenase dye terminator cycle sequencing premix. SAC1cDNA was a 2616 nucleotide encoding 871 amino acids. The sequences are shown as SEQ ID No: 1 (amino acid sequence) and SEQ ID No: 2 (nucleotide sequence) in Sequence Listing. The plasmid in which cDNA of SAC1 is inserted was designated pmSAC1 TOPO and originally deposited with the Biotechnology Laboratory, National Institute of Advanced Science and Technology, the Ministry of Economy, Trade and Industry (1-3, Higashi 1-Chome, Tsukuba-shi, Ibaraki-ken, Japan 305-8566) on Nov. 30, 1999. The deposition was subject to a request to transfer to a deposition under the Budapest Treaty on Nov. 1, 2000, and received as FERM BP-7345.

[0030] Next, expression of SAC1 in each tissue was confirmed by Northern hybridization (65° C.) using SAC1 cDNA labeled with ³²P as a probe. As a result, SAC1 mRNA with a length of about 3 Kb was confirmed to have been expressed only in the kidney (FIG. 1(a)).

[0031] In addition, to isolate cDNA encoding the human origin SAC (hSAC), a human kidney cDNA library was screened using ³²P-labeled SAC1 cDNA as a probe. The resulting clone was subcloned into vectors M13 mp1, M13 mp19 (Takara, Ltd.) and pGEM3Z (Promega Co.) to determine the nucleotide sequence in the same manner as above. The sequences are shown in SEQ ID No: 3 (amino acid sequence) and SEQ ID No: 4 (nucleotide sequence) in Sequence-Listing.

Example 2

[0032] Antibody Preparation and Immunostaining

[0033] An antibody to the specific peptide in the C-terminal (844-853) of SAC1 protein (hereinafter referred to as “C-terminal peptide”) (SEQ ID No: 10 in Sequence Listing) bound to polyethylene glycol was prepared. The C-terminal peptide of the mouse SAC1 protein was bound to the polyethylene glycol (PEG) in accordance with the method of Maeda et al. (Biochem. Biophys. Res. Comm. (1998) 248. 485-489). Briefly, one equivalent dicyclohexylcarbodiimide was added to oxidized PEG dissolved in dichloromethane (Wako Pure Chemicals Co., Ltd.) and the mixture was stirred at 0° C. Then, 10 equivalents of ethylenediamine were added and the mixture was stirred overnight. The product obtained after evaporating the solvent from the filtrate was dissolved in 50% acetic acid. Amino acid-type PEG (aaPEG) was purified by a Sephadex G-25 column and the N-terminal amino group was converted into an Fmoc group to prepare Fmoc-aaPEG. Fmoc-aaPEG and Boc-PLD(Pmc)NLGNPNC-OH were bound to a Rink resin (300 mg; Watanabe Chemical Co., Ltd.) in that order using the diisopropyl carbodiimide/1-hydroxybenzotriazole method (Konig and Geiger, Chem. Ber. (1970) 103, 788-798,2024-2033). After removing the Fmoc group, the resin was treated with trifluoroacetic acid/thioanisole/anisole/ethanedithiol (94/2/2/2) for 3 hours to cleave out C-terminal peptide-aaPEG. The crude peptide precipitated by adding ether to the filtrate was purified using reverse phase HPLC, thereby obtaining 43 mg of C-terminal peptide-aaPEG.

[0034] Two NZW rabbits were immunized by intramuscular injection of 1 mg of the antigen (C-terminal peptide-aaPEG) emulsified with Freund's complete adjuvant. Thereafter, the rabbits were immunized in the same manner every two weeks using the same amount of antigen emulsified with Freund's incomplete adjuvant. During 4-8 weeks after the administration, the antibody titer of serum was measured by an ELISA method. The serum exhibiting titer of 10,000 times or more than that of the control serum was prepared. The antibody was purified from the serum by Protein A column (Pharmacia Co.), followed by affinity purification using a Proton kit (Multiple Peptide System Co.). After staining a fresh kidney slice with the antibody diluted to 500 fold, the protein was detected using an anti-rabbit IgG antibody (DAKO Co.) labeled with FITC. As a result, the basolateral membrane of proximal tubule was stained, it was thus confirmed that the SAC1 protein is localized (see FIG. 1(b))

Example 3

[0035] Expression of cDNA

[0036] The plasmid (pEGFP-N1; Clonetech Co.) expressing a green fluorescent protein was used as a marker for transfection. For the transient expression of protein, CHO cells were cultured in a HamF-12 culture medium (Gibco-BRL Co.) supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin. On the day before transformation, CHO cells (10⁵ cells) were seeded to a 35 mm dish in which a cover slip coated with rat tail collagen was placed. On the next day, SAC1 plasmid (1 μg) in PCMV-SPORT (Gibco-BRL Co.) and pEGFP-N1 (1 μg) were added per one transfection to a mixture of serum-free medium (97 μl) and FuGENE6 (Roche, 3 μl) which had been incubated for 5 minutes at room temperature. After incubation for 15 minutes at room temperature, the mixture was added to a dish containing 3 ml of serum-added medium. Cells grown on the cover slip were used as a patch clamp sample. A 10% FCS-containing culture medium was used for the growth and transfection. The electrophysiological test was started after 24 hours and the fluorescence measurement was carried out 48 hours after the transfection.

Example 4 Fluorescence Measurement of SAC1 Expression Cells

[0037] For 2-3 hours prior to start of the test, the transformed cells were incubated with 10 μM fura-2AM (Dojin Co.) dissolved in a serum-free culture medium. GFP fluorescence at 480 nm and fura-2 fluorescence at 360/380 nm were visualized by a manual exchange of dichroic Olympus system mirrors (Merlin Co.). Images for the fluorescence at 480 nm and fluorescence at 360/380 nm were obtained every two seconds. The fluorescence intensity of the objective area was calculated for analysis. Cells were bathed in a solution of 125 mM NaCl, 5 mM KCl, 1.2 mM MgSO₄, 1 mM Na₂HPO₄, 1 mM CaCl₂, and 3 mM HEPES (pH 7.4) at 34° C. To produce a mechanical stress directly to cells, they were touched by a glass pipette with a round tip using an electrically controlled micromanipulator (Model 5170; Eppendorf Co.). The results obtained by inducing a mechanical stimulus to light cells (GFP positive) using the glass pipette are shown in FIG. 2 and Table 1. The manifest change of the fura-2 fluorescence ratio in the SAC1 expression cells as compared with the control cells was observed, thus it was indicated that the proteins of the present invention function as channel proteins, clearly bringing about an alteration of [Ca²⁺]i (intracellular calcium ion concentration). TABLE 1 no added Gd³⁺ added Gd³⁺ Control cells  4.5 ± 3.8 (8) SAC1 expression cells 12.8 ± 1.4** (8) 4.8 ± 1.4 (10)

Example 5

[0038] Single Channel Analysis of SAC1 Expression Cells

[0039] Patch clamp recordings were carried out according to the method described in a previous paper (Suzuki, M., Sato, J., Kutsuwada, K., Ooki, G. and Imai, M. (1999) J. Biol. Chem. 274, 6330-5). GFP positive cells were visualized by a fluorescence measurement with an emission of 490 nm (CAM 2000 system; Jusco Co., Ltd.). Cells were set on a mount under a vectorial flow of solution at a rate of 1 ml/ml. To control the temperature, the solution was heated to 34° C. by a DC supply (Model LPD; Kikusui Electronic Co., Ltd.) prior to mounting. A solution of 140 mM NaCl, 5 mM KCl, 1.2 mM MgSO₄, 1 mM Na₂HPO₄, 1 mM CaCl₂, and 3 mM HEPES was used as a bath solution. In single channel analysis, the current was recorded with an EPC-7 patch clamp amplifier (List-Electronic Co., Ltd.) and stored on a DAT recorder (DAT-200; Sony Corp.) at 10 KHz. A 5 KHz filter was applied for the analysis. The records were sampled by Fetchex (Software Axon, version 6.0) to analyze the open probability. The data were analyzed using Igor ver. 2.01 and Patch Analysist ver. 1.21. A digital filter was used at 2 KHz to calculate the open probability by current distribution analysis. To analyze the open probability in the presence of GdCl₃, single channel amplitude was set for those without reagent and the mean open probability (Po) was determined by the records during 10 seconds. The pipette pressure was adjusted using a manometer and negative pressure was manually changed. To examine the channel opening, negative pressure was applied through a cell attaching pipette using a 150 mM NaCl solution. As a result, more than one half of the patches on expressed cells revealed channel activity by negative pressure. As shown in FIG. 3A, the SAC1 channel abruptly opens at 30 mmHg negative pressure. The open probability suddenly reached the state of full opening. In order to confirm this phenomenon, the suction was gradually applied (FIG. 3B). The channel suddenly opened as if there was a threshold for opening. The threshold was constant (33+2.3 mmHg) and reproducible. Current-voltage relationship was measured with pipettes filled with various solutions (sodium chloride, sodium gluconate and potassium chloride) to confirm that there was observed no significant changes. A single channel conductance was measured as slope conductance with cell-attach configuration. When sodium chloride was substituted with sodium gluconate or potassium chloride, there was almost no change in the parameters, thus it was clearly confirmed that the channel is non-selective with respect to cations. Accumulation of all-or-none openings of individual channels with different sensitivities to pressure, resulted in an S-shaped opening response to stretch. The mean open probability (Po) however did not fit to the Bolzmann's function of the pressure. The response was not affected when 0.5 μM GdCl₃ was added to the pipette, confirming that the protein possesses a function as a mechanosensitive channel protein (FIG. 3C)

INDUSTRIAL APPLICABILITY

[0040] According to the present invention, novel mechanosensitive channel protein derived from mice or humans (SAC1 (mouse) and hSAC (human)) are provided. The proteins possess a function of being expressed specifically in the kidney and non-selectively incorporating cations into cells in response to mechanical stimulus. DNA encoding these proteins, a method of screening cation channel activators or inhibitors using said proteins and antibodies to the proteins are also provided.

[0041] The proteins, DNA, and antibodies to the proteins can be used as a diagnostic agent or a therapeutic agent for diseases based on abnormalities in the cation channel such as hypertension or diabetes or as a tool for screening chemicals for preventing and/or treating these diseases.

[0042] Remarks to Deposited Biological Materials

[0043] a. Name and address of the organization in which the biological materials have been deposited:

[0044] Name: The Biotechnology Laboratory, National Institute of Advanced Science and Technology, the Ministry of Economy, Trade and Industry

[0045] Address: 1-1-3, Higashi, Tsukuba-shi, Ibaraki-ken, Japan (Postal Code: 305-3566).

[0046] b. Date of deposition: Nov. 30, 1999 (Original deposition)

[0047] c. Number of deposition given by the deposition organization:

[0048] FERM BP-7345

1 10 1 871 PRT Mus musculus 1 Met Ala Asp Pro Gly Asp Gly Pro Arg Ala Ala Pro Gly Glu Val Ala 1 5 10 15 Glu Pro Pro Gly Asp Glu Ser Gly Thr Ser Gly Gly Glu Ala Phe Pro 20 25 30 Leu Ser Ser Leu Ala Asn Leu Phe Glu Gly Glu Glu Gly Ser Ser Ser 35 40 45 Leu Ser Pro Val Asp Ala Ser Arg Pro Ala Gly Pro Gly Asp Gly Arg 50 55 60 Pro Asn Leu Arg Met Lys Phe Gln Gly Ala Phe Arg Lys Gly Val Pro 65 70 75 80 Asn Pro Ile Asp Leu Leu Glu Ser Thr Leu Tyr Glu Ser Ser Val Val 85 90 95 Pro Gly Pro Lys Lys Ala Pro Met Asp Ser Leu Phe Asp Tyr Gly Thr 100 105 110 Tyr Arg His His Pro Ser Asp Asn Lys Arg Trp Arg Arg Lys Val Val 115 120 125 Glu Lys Gln Pro Gln Ser Pro Lys Thr Pro Ala Pro Gln Pro Pro Pro 130 135 140 Ile Leu Lys Val Phe Asn Arg Pro Ile Leu Phe Asp Ile Val Ser Arg 145 150 155 160 Gly Ser Thr Ala Asp Leu Asp Gly Leu Leu Ser Phe Leu Leu Thr His 165 170 175 Lys Lys Arg Leu Thr Asp Glu Glu Phe Arg Glu Pro Ser Thr Gly Lys 180 185 190 Thr Cys Leu Pro Lys Ala Leu Leu Asn Leu Ser Asn Gly Arg Asn Asp 195 200 205 Thr Leu Gln Val Leu Leu Asp Ile Ala Glu Arg Thr Gly Asn Met Arg 210 215 220 Glu Phe Ile Asn Ser Pro Phe Arg Asp Ile Tyr Tyr Arg Gly Gln Thr 225 230 235 240 Ser Leu His Ile Ala Ile Glu Arg Arg Cys Lys His Tyr Val Glu Leu 245 250 255 Leu Val Ala Gln Gly Ala Asp Val His Ala Gln Ala Arg Gly Arg Phe 260 265 270 Phe Gln Pro Lys Asp Glu Gly Gly Tyr Phe Tyr Phe Gly Glu Leu Pro 275 280 285 Leu Ser Leu Ala Ala Cys Thr Asn Gln Pro His Ile Val Asn Tyr Leu 290 295 300 Thr Glu Asn Pro His Lys Lys Ala Asp Met Arg Arg Gln Asp Ser Arg 305 310 315 320 Gly Asn Thr Val Leu His Ala Leu Val Ala Ile Ala Asp Asn Thr Arg 325 330 335 Glu Asn Thr Lys Phe Val Thr Lys Met Tyr Asp Leu Leu Leu Leu Lys 340 345 350 Cys Ser Arg Leu Phe Pro Asp Ser Asn Leu Glu Thr Val Leu Asn Asn 355 360 365 Asp Gly Leu Ser Pro Leu Met Met Ala Ala Lys Thr Gly Lys Ile Gly 370 375 380 Val Phe Gln His Ile Ile Arg Arg Glu Val Thr Asp Glu Asp Thr Arg 385 390 395 400 His Leu Ser Arg Lys Phe Lys Asp Trp Ala Tyr Gly Pro Val Tyr Ser 405 410 415 Ser Leu Tyr Asp Leu Ser Ser Leu Asp Thr Cys Gly Glu Glu Val Ser 420 425 430 Val Leu Glu Ile Leu Val Tyr Asn Ser Lys Ile Glu Asn Arg His Glu 435 440 445 Met Leu Ala Val Glu Pro Ile Asn Glu Leu Leu Arg Asp Lys Trp Arg 450 455 460 Lys Phe Gly Ala Val Ser Phe Tyr Ile Asn Val Val Ser Tyr Leu Cys 465 470 475 480 Ala Met Val Ile Phe Thr Leu Thr Ala Tyr Tyr Gln Pro Leu Glu Gly 485 490 495 Thr Pro Pro Tyr Pro Tyr Arg Thr Thr Val Asp Tyr Leu Arg Leu Ala 500 505 510 Gly Glu Val Ile Thr Leu Phe Thr Gly Val Leu Phe Phe Phe Thr Ser 515 520 525 Ile Lys Asp Leu Phe Thr Lys Lys Cys Pro Gly Val Asn Ser Leu Phe 530 535 540 Val Asp Gly Ser Phe Gln Leu Leu Tyr Phe Ile Tyr Ser Val Leu Val 545 550 555 560 Val Val Ser Ala Ala Leu Tyr Leu Ala Gly Ile Glu Ala Tyr Leu Ala 565 570 575 Val Met Val Phe Ala Leu Val Leu Gly Trp Met Asn Ala Leu Tyr Phe 580 585 590 Thr Arg Gly Leu Lys Leu Thr Gly Thr Tyr Ser Ile Met Ile Gln Lys 595 600 605 Ile Leu Phe Lys Asp Leu Phe Arg Phe Leu Leu Val Tyr Leu Leu Phe 610 615 620 Met Ile Gly Tyr Ala Ser Ala Leu Val Thr Leu Leu Asn Pro Cys Thr 625 630 635 640 Asn Met Lys Val Cys Asp Glu Asp Gln Ser Asn Cys Thr Val Pro Thr 645 650 655 Tyr Pro Ala Cys Arg Asp Ser Glu Thr Phe Ser Ala Phe Leu Leu Asp 660 665 670 Leu Phe Lys Leu Thr Ile Gly Met Gly Asp Leu Glu Met Leu Ser Ser 675 680 685 Ala Lys Tyr Pro Val Val Phe Ile Leu Leu Leu Val Thr Tyr Ile Ile 690 695 700 Leu Thr Phe Val Leu Leu Leu Asn Met Leu Ile Ala Leu Met Gly Glu 705 710 715 720 Thr Val Gly Gln Val Ser Lys Glu Ser Lys His Ile Trp Lys Leu Gln 725 730 735 Trp Ala Thr Thr Ile Leu Asp Ile Glu Arg Ser Phe Pro Val Phe Leu 740 745 750 Arg Lys Ala Phe Arg Ser Gly Glu Met Val Thr Val Gly Lys Ser Ser 755 760 765 Asp Gly Thr Pro Asp Arg Arg Trp Cys Phe Arg Val Asp Glu Val Ser 770 775 780 Trp Ser His Trp Asn Gln Asn Leu Gly Ile Ile Asn Glu Asp Pro Gly 785 790 795 800 Lys Ser Glu Ile Tyr Gln Tyr Tyr Gly Phe Ser His Thr Val Gly Arg 805 810 815 Leu Arg Arg Asp Arg Trp Ser Ser Val Val Pro Arg Val Val Glu Leu 820 825 830 Asn Lys Asn Ser Ser Ala Asp Glu Val Val Val Pro Leu Asp Asn Leu 835 840 845 Gly Asn Pro Asn Cys Asp Gly His Gln Gln Gly Tyr Ala Pro Lys Trp 850 855 860 Arg Thr Asp Asp Ala Pro Leu 865 870 2 2616 DNA Mus musculus 2 atggcagatc ctggtgatgg tccccgtgca gcgcctgggg aggtggctga gccccctgga 60 gatgagagtg gtacctctgg tggggaggcc ttccccctct cttccctggc caatctgttt 120 gagggggagg aaggctcctc ttctctttcc ccggtggatg ctagccgccc tgctggccct 180 ggcgatggac gtccaaacct gcgtatgaag ttccagggcg ctttccgcaa gggggttccc 240 aaccccattg acctgttgga gtccaccctg tacgagtcct cagtagtgcc tgggcccaag 300 aaagcgccca tggattcctt gttcgactac ggcacttacc gtcaccaccc cagtgacaac 360 aagagatgga ggagaaaggt cgtggagaag cagccacaga gccccaaaac tcctgcaccc 420 cagccacccc ccatcctcaa agtcttcaat cggcccatcc tctttgacat tgtgtcccgg 480 ggctccactg cggacctaga tggactgctc tccttcttgt tgacccacaa gaagcgcctg 540 actgatgagg agttccggga gccgtccacg gggaagacct gcctgcccaa ggcgctgctg 600 aacctaagca acgggcgcaa cgacaccctc caggtgttgc tggacattgc ggagcgcacc 660 ggcaacatgc gtgaattcat caactcgccc ttcagagaca tctactaccg aggccagaca 720 tccctgcaca ttgccatcga acggcgctgc aagcactacg tggagctgct ggtggcccag 780 ggagccgacg tgcacgccca ggcccgcggc cgcttcttcc agcccaagga tgagggaggc 840 tacttctact ttggggagct gcccttgtcc ctggcagcct gcaccaacca gccgcacatc 900 gtcaactacc tgacagagaa ccctcacaag aaagctgaca tgaggcgaca ggactcgagg 960 gggaacacgg tgctgcacgc gctggtggcc atcgccgaca acacccgaga gaacaccaag 1020 tttgtcacca agatgtacga cctgctgctt ctcaagtgtt cacgcctctt ccccgacagc 1080 aacctggaga cagttctcaa caatgatggc ctttcgcctc tcatgatggc tgccaagaca 1140 ggcaagatcg gggtctttca gcacatcatc cgacgtgagg tgacagatga ggacacccgg 1200 catctgtctc gcaagttcaa ggactgggcc tatgggcctg tgtattcttc tctctacgac 1260 ctctcctccc tggacacatg cggggaggag gtgtccgtgc tggagatcct ggtgtacaac 1320 agcaagatcg agaaccgcca tgagatgctg gctgtagagc ccattaacga actgttgaga 1380 gacaagtggc gtaagtttgg ggctgtgtcc ttctacatca acgtggtctc ctatctgtgt 1440 gccatggtca tcttcaccct caccgcctac tatcagccac tggagggcac gccaccctac 1500 ccttaccgga ccacagtgga ctacctgagg ctggctggcg aggtcatcac gctcttcaca 1560 ggagtcctgt tcttctttac cagtatcaaa gacttgttca cgaagaaatg ccctggagtg 1620 aattctctct tcgtcgatgg ctccttccag ttactctact tcatctactc tgtgctggtg 1680 gttgtctctg cggcgctcta cctggctggg atcgaggcct acctggctgt gatggtcttt 1740 gccctggtcc tgggctggat gaatgcgctg tacttcacgc gcgggttgaa gctgacgggg 1800 acctacagca tcatgattca gaagatcctc ttcaaagacc tcttccgctt cctgcttgtg 1860 tacctgctct tcatgatcgg ctatgcctca gccctggtca ccctcctgaa tccgtgcacc 1920 aacatgaagg tctgtgacga ggaccagagc aactgcacgg tgcccacgta tcctgcgtgc 1980 cgcgacagcg agaccttcag cgccttcctc ctggacctct tcaagctcac catcggcatg 2040 ggagacctgg agatgctgag cagcgccaag taccccgtgg tcttcatcct cctgctggtc 2100 acctacatca tcctcacctt cgtgctcctg ttgaacatgc ttatcgccct catgggtgag 2160 accgtgggcc aggtgtccaa ggagagcaag cacatctgga agttgcagtg ggccaccacc 2220 atcctggaca tcgagcgttc cttccctgtg ttcctgagga aggccttccg ctccggagag 2280 atggtgactg tgggcaagag ctcagatggc actccggacc gcaggtggtg cttcagggtg 2340 gacgaggtga gctggtctca ctggaaccag aacttgggca tcattaacga ggaccctggc 2400 aagagtgaaa tctaccagta ctatggcttc tcccacaccg tggggcgcct tcgtagggat 2460 cgttggtcct cggtggtgcc ccgcgtagtg gagctgaaca agaactcaag cgcagatgaa 2520 gtggtggtac ccctggataa cctagggaac cccaactgtg acggccacca gcagggctac 2580 gctcccaagt ggaggacgga cgatgcccca ctgtag 2616 3 871 PRT Homo sapiens 3 Met Ala Asp Ser Ser Glu Gly Pro Arg Ala Gly Pro Gly Glu Val Ala 1 5 10 15 Glu Leu Pro Gly Asp Glu Ser Gly Thr Pro Gly Gly Glu Ala Phe Pro 20 25 30 Leu Ser Ser Leu Ala Asn Leu Phe Glu Gly Glu Asp Gly Ser Leu Ser 35 40 45 Pro Ser Pro Ala Asp Ala Ser Arg Pro Ala Gly Pro Gly Asp Gly Arg 50 55 60 Pro Asn Leu Arg Met Lys Phe Gln Gly Ala Phe Arg Lys Gly Val Pro 65 70 75 80 Asn Pro Ile Asp Leu Leu Glu Ser Thr Leu Tyr Glu Ser Ser Val Val 85 90 95 Pro Gly Pro Lys Lys Ala Pro Met Asp Ser Leu Phe Asp Tyr Gly Thr 100 105 110 Tyr Arg His His Ser Ser Asp Asn Lys Arg Trp Arg Lys Lys Ile Ile 115 120 125 Glu Lys Gln Pro Gln Ser Pro Lys Ala Pro Ala Pro Gln Pro Pro Pro 130 135 140 Ile Leu Lys Val Phe Asn Arg Pro Ile Leu Phe Asp Ile Val Ser Arg 145 150 155 160 Gly Ser Thr Ala Asp Leu Asp Gly Leu Leu Pro Phe Leu Leu Thr His 165 170 175 Lys Lys Arg Leu Thr Asp Glu Glu Phe Arg Glu Pro Ser Thr Gly Lys 180 185 190 Thr Cys Leu Pro Lys Ala Leu Leu Asn Leu Ser Asn Gly Arg Asn Asp 195 200 205 Thr Ile Pro Val Leu Leu Asp Ile Ala Glu Arg Thr Gly Asn Met Arg 210 215 220 Glu Phe Ile Asn Ser Pro Phe Arg Asp Ile Tyr Tyr Arg Gly Gln Thr 225 230 235 240 Ala Leu His Ile Ala Ile Glu Arg Arg Cys Lys His Tyr Val Glu Leu 245 250 255 Leu Val Ala Gln Gly Ala Asp Val His Ala Gln Ala Arg Gly Arg Phe 260 265 270 Phe Gln Pro Lys Asp Glu Gly Gly Tyr Phe Tyr Phe Gly Glu Leu Pro 275 280 285 Leu Ser Leu Ala Ala Cys Thr Asn Gln Pro His Ile Val Asn Tyr Leu 290 295 300 Thr Glu Asn Pro His Lys Lys Ala Asp Met Arg Arg Gln Asp Ser Arg 305 310 315 320 Gly Asn Thr Val Leu His Ala Leu Val Ala Ile Ala Asp Asn Thr Arg 325 330 335 Glu Asn Thr Lys Phe Val Thr Lys Met Tyr Asp Leu Leu Leu Leu Lys 340 345 350 Cys Ala Arg Leu Phe Pro Asp Ser Asn Leu Glu Ala Val Leu Asn Asn 355 360 365 Asp Gly Leu Ser Pro Leu Met Met Ala Ala Lys Thr Gly Lys Ile Gly 370 375 380 Val Phe Gln His Ile Ile Arg Arg Glu Val Thr Asp Glu Asp Thr Arg 385 390 395 400 His Leu Ser Arg Lys Phe Lys Asp Trp Ala Tyr Gly Pro Val Tyr Ser 405 410 415 Ser Leu Tyr Asp Leu Ser Ser Leu Asp Thr Cys Gly Glu Glu Ala Ser 420 425 430 Val Leu Glu Ile Leu Val Tyr Asn Ser Lys Ile Glu Asn Arg His Glu 435 440 445 Met Leu Ala Val Glu Pro Ile Asn Glu Leu Leu Arg Asp Lys Trp Arg 450 455 460 Lys Phe Gly Ala Val Ser Phe Tyr Ile Asn Val Val Ser Tyr Leu Cys 465 470 475 480 Ala Met Val Ile Phe Thr Leu Thr Ala Tyr Tyr Gln Pro Leu Glu Gly 485 490 495 Thr Val Pro Tyr Pro Tyr Arg Thr Thr Val Asp Tyr Leu Arg Leu Ala 500 505 510 Gly Glu Val Ile Thr Leu Phe Thr Gly Val Leu Phe Phe Phe Thr Asn 515 520 525 Ile Lys Asp Leu Phe Met Lys Lys Cys Pro Gly Val Asn Ser Leu Phe 530 535 540 Ile Asp Gly Ser Phe Gln Leu Leu Ser Phe Ile Tyr Ser Val Leu Val 545 550 555 560 Ile Val Ser Ala Ala Leu Tyr Leu Ala Gly Ile Glu Ala Tyr Leu Ala 565 570 575 Val Met Val Phe Ala Leu Val Leu Gly Trp Met Asn Ala Leu Tyr Phe 580 585 590 Thr Arg Gly Leu Lys Leu Thr Gly Thr Tyr Ser Ile Met Ile Gln Lys 595 600 605 Val Leu Phe Lys Asp Leu Phe Arg Phe Leu Leu Val Tyr Leu Leu Phe 610 615 620 Met Ile Gly Tyr Ala Ser Ala Leu Val Ser Leu Leu Asn Pro Cys Ala 625 630 635 640 Asn Met Lys Val Cys Asn Glu Asp Gln Thr Asn Cys Thr Val Pro Thr 645 650 655 Tyr Pro Ser Cys Arg Asp Ser Glu Thr Phe Ser Thr Phe Leu Leu Asp 660 665 670 Leu Phe Lys Leu Thr Ile Gly Met Gly Asp Leu Glu Met Leu Ser Ser 675 680 685 Thr Lys Tyr Pro Val Val Phe Ile Ile Leu Leu Val Thr Tyr Ile Ile 690 695 700 Leu Thr Phe Val Leu Leu Leu Asn Met Leu Ile Ala Leu Met Gly Glu 705 710 715 720 Thr Val Gly Gln Val Ser Lys Glu Ser Lys His Ile Trp Lys Leu Gln 725 730 735 Trp Ala Thr Thr Ile Leu Asp Ile Glu Arg Ser Phe Pro Val Phe Leu 740 745 750 Arg Lys Ala Phe Arg Ser Gly Glu Met Val Thr Val Gly Lys Ser Ser 755 760 765 Asp Gly Thr Pro Asp Arg Arg Trp Cys Phe Arg Val Asn Glu Val Asn 770 775 780 Trp Ser His Trp Asn Gln Asn Leu Gly Ile Ile Asn Glu Asp Pro Gly 785 790 795 800 Lys Asn Glu Thr Tyr Gln Tyr Tyr Gly Phe Ser His Thr Val Gly Arg 805 810 815 Leu Arg Met Asp Arg Trp Ser Ser Val Val Pro Arg Val Val Glu Leu 820 825 830 Asn Lys Asn Ser Asn Pro Asp Glu Val Val Val Pro Leu Asp Ser Met 835 840 845 Gly Asn Pro Arg Cys Asp Gly His Gln Gln Gly Tyr Pro Arg Lys Trp 850 855 860 Arg Thr Asp Asp Ala Pro Leu 865 870 4 2616 DNA Homo sapiens 4 atggcggatt ccagcgaagg cccccgcgcg gggcccgggg aggtggctga gctccccggg 60 gatgagagtg gcaccccagg tggggaggct tttcctctct cctccctggc caatctgttt 120 gagggggagg atggctccct ttcgccctca ccggctgatg ccagtcgccc tgctggccca 180 ggcgatgggc gaccaaatct gcgcatgaag ttccagggcg ccttccgcaa gggggtgccc 240 aaccccatcg atctgctgga gtccacccta tatgagtcct cggtggtgcc tgggcccaag 300 aaagcaccca tggactcact gtttgactac ggcacctatc gtcaccactc cagtgacaac 360 aagaggtgga ggaagaagat catagagaag cagccgcaga gccccaaagc ccctgcccct 420 cagccgcccc ccatcctcaa agtcttcaac cggcctatcc tctttgacat cgtgtcccgg 480 ggctccactg ctgacctgga cgggctgctc ccattcttgc tgacccacaa gaaacgccta 540 actgatgagg agtttagaga gccatctacg gggaagacct gcctgcccaa ggccttgctg 600 aacctgagca atggccgcaa cgacaccatc cctgtgctgc tggacatcgc ggagcgcacc 660 ggcaacatga gggagttcat taactcgccc ttccgtgaca tctactatcg aggtcagaca 720 gccctgcaca tcgccattga gcgtcgctgc aaacactacg tggaacttct cgtggcccag 780 ggagctgatg tccacgccca ggcccgtggg cgcttcttcc agcccaagga tgaggggggc 840 tacttctact ttggtgagct gcccctgtcg ctggctgcct gcaccaacca gccccacatt 900 gtcaactacc tgacggagaa cccccacaag aaggcggaca tgcggcgcca ggactcgcga 960 ggcaacacag tgctgcatgc gctggtggcc attgctgaca acacccgtga gaacaccaag 1020 tttgttacca agatgtacga cctgctgctg ctcaagtgtg cccgcctctt ccccgacagc 1080 aacctggagg ccgtgctcaa caacgacggc ctctcgcccc tcatgatggc tgccaagacg 1140 ggcaagattg gggtgtttca gcacatcatc cggcgggagg tgacggatga ggacacacgg 1200 cacctgtccc gcaagttcaa ggactgggcc tatgggccag tgtattcctc gctttatgac 1260 ctctcctccc tggacacgtg tggggaagag gcctccgtgc tggagatcct ggtgtacaac 1320 agcaagattg agaaccgcca cgagatgctg gctgtggagc ccatcaatga actgctgcgg 1380 gacaagtggc gcaagttcgg ggccgtctcc ttctacatca acgtggtctc ctacctgtgt 1440 gccatggtca tcttcactct caccgcctac taccagccgc tggagggcac agtgccgtac 1500 ccttaccgca ccacggtgga ctacctgcgg ctggctggcg aggtcattac gctcttcact 1560 ggggtcctgt tcttcttcac caacatcaaa gacttgttca tgaagaaatg ccctggagtg 1620 aattctctct tcattgatgg ctccttccag ctgctcagct tcatctactc tgtcctggtg 1680 atcgtctcag cagccctcta cctggcaggg atcgaggcct acctggccgt gatggtcttt 1740 gccctggtcc tgggctggat gaatgccctt tacttcaccc gtgggctgaa gctgacgggg 1800 acctatagca tcatgatcca gaaggtactc ttcaaggacc ttttccgatt cctgctcgtc 1860 tacttgctct tcatgatcgg ctacgcttca gccctggtct ccctcctgaa cccgtgtgcc 1920 aacatgaagg tgtgcaatga ggaccagacc aactgcacag tgcccactta cccctcgtgc 1980 cgtgacagcg agaccttcag caccttcctc ctggacctgt ttaagctgac catcggcatg 2040 ggcgacctgg agatgctgag cagcaccaag taccccgtgg tcttcatcat cctgctggtg 2100 acctacatca tcctcacctt tgtgctgctc ctcaacatgc tcattgccct catgggcgag 2160 acagtgggcc aggtctccaa ggagagcaag cacatctgga agctgcagtg ggccaccacc 2220 atcctggaca ttgagcgctc cttccccgta ttcctgagga aggccttccg ctctggggag 2280 atggtcaccg tgggcaagag ctcggacggc actcctgacc gcaggtggtg cttcagggtg 2340 aatgaggtga actggtctca ctggaaccag aacttgggca tcatcaacga ggacccgggc 2400 aagaatgaga cctaccagta ttatggcttc tcgcataccg tgggccgcct ccgaatggat 2460 cgctggtcct cggtggtacc ccgcgtggtg gaactgaaca agaactcgaa cccggacgag 2520 gtggtggtgc ctctggacag catggggaac ccccgctgcg atggccacca gcagggttac 2580 ccccgcaagt ggaggactga tgacgccccg ctctag 2616 5 19 DNA Artificial sequence Synthesized DNA 5 gagagaacac caagtttgt 19 6 26 DNA Artificial sequence Synthesized DNA 6 cagcggcccc aatctcttca aagtac 26 7 20 DNA Artificial sequence Synthesized DNA 7 ctacagccag catctcatgg 20 8 30 DNA Artificial sequence Synthesized DNA 8 tcagggtcca gtatggcaga tcctggtgat 30 9 22 DNA Artificial sequence Synthesized DNA 9 gttaatgggc tctacagcca gc 22 10 10 PRT Homo sapiens 10 Pro Leu Asp Asn Leu Gly Asn Pro Asn Cys 1 5 10 

1. A protein wherein said protein is expressed specifically in the kidney and has a function of transporting cations non-selectively into cells in response to a mechanical stimulus.
 2. The protein according to claim 1, which is derived from mice.
 3. The protein according to claim 2, having an amino acid sequence shown in SEQ ID No: 1 in the Sequence Listing.
 4. A DNA encoding the amino acid sequence described in claim
 3. 5. The DNA according to claim 4, having a nucleotide sequence shown in SEQ ID No: 2 in the Sequence Listing.
 6. The protein according to claim 1, which is derived from humans.
 7. The protein according to claim 6, having an amino acid sequence shown in SEQ ID No: 3 in the Sequence Listing.
 8. A DNA encoding the amino acid sequence described in claim
 7. 9. The DNA according to claim 8, having a nucleotide sequence shown in SEQ ID No: 4 in the Sequence Listing.
 10. A protein according to claim 3 or 7, having an amino acid sequence in which one or more amino acids are deleted from, substituted in or added to the given sequence.
 11. A DNA hybridizing with the DNA described in any one of claims 4, 5, 8, and
 9. 12. A method of screening cation channel activators or inhibitors using the protein described in any one of claims 1, 2, 3, 6, and
 7. 13. An antibody to the protein described in any one of claims 1, 2, 3, 6, and
 7. 14. The antibody according to claim 13, wherein the antibody is a monoclonal antibody. 